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Titolo:
Comparison of several techniques for the detection of apoptotic astrocytesin vitro
Autore:
Micoud, F; Mandrand, B; Malcus-Vocanson, C;
Indirizzi:
Ecole Normale Super Lyon, CNRS BioMerieux, Unite Mixte 103, F-69364 Lyon, France Ecole Normale Super Lyon Lyon France F-69364 e 103, F-69364 Lyon, France
Titolo Testata:
CELL PROLIFERATION
fascicolo: 2, volume: 34, anno: 2001,
pagine: 99 - 113
SICI:
0960-7722(200104)34:2<99:COSTFT>2.0.ZU;2-1
Fonte:
ISI
Lingua:
ENG
Soggetto:
MULTIPLE-SCLEROSIS BRAIN; PROGRAMMED CELL-DEATH; END LABELING TUNEL; DNA FRAGMENTATION; ADHERENT CELLS; THYMOCYTE APOPTOSIS; FLOW-CYTOMETRY; NERVOUS-SYSTEM; ANNEXIN-V; PHOSPHATIDYLSERINE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
53
Recensione:
Indirizzi per estratti:
Indirizzo: Malcus-Vocanson, C BioMerieux, Chemin Orme, F-69280 Marcy Etoile, France BioMerieux Chemin Orme Marcy Etoile France F-69280 nce
Citazione:
F. Micoud et al., "Comparison of several techniques for the detection of apoptotic astrocytesin vitro", CELL PROLIF, 34(2), 2001, pp. 99-113

Abstract

Implication of apoptosis in numerous physiological and pathological processes has resulted in the development of numerous methods to detect apoptosis, but none of them is adapted to all cell types. In this study, we induced apoptosis on murine immortalized astrocytes with urine from multiple sclerosis (MS) patients. Among techniques allowing the detection of apoptotic cells, only a few are adapted to adherent cells such as astrocytes We comparedseveral techniques (propidium iodide labelling and flow cytometry analysis, TUNEL and annexin V labelling in immunofluorescence, DNA ladder, ELISA tests to detect nucleosomes) in order to choose the method best adapted to our adherent cellular model and to discuss their practicability for the detection of apoptosis on adherent cells. For technical course, propidium iodide labelling followed by flow cytometry analysis as a quantitative technique, and TUNEL in IF (easier and quickerthan propidium iodide) as a semiquantitative test were both retained as best adapted to our case. Moreover, in our model, we have observed that phosphatydilserine externalization and DNA fragmentation were concomittant after induction of apoptosis. Techniques studied in this article would allow an enlarged study of the apoptotic mechanism in several pathologies by culture of adherent cells sensitive to apoptosis in vitro.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 10/07/20 alle ore 10:44:32